Immune Netw.  2012 Feb;12(1):8-17. 10.4110/in.2012.12.1.8.

Baculovirus-based Vaccine Displaying Respiratory Syncytial Virus Glycoprotein Induces Protective Immunity against RSV Infection without Vaccine-Enhanced Disease

• Kim S
• Chang J

• KMID: 2014651
• DOI: http://doi.org/10.4110/in.2012.12.1.8

Abstract

BACKGROUND
Respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract diseases in infancy and early childhood. Despite its importance as a pathogen, there is no licensed vaccine against RSV yet. The attachment glycoprotein (G) of RSV is a potentially important target for protective antiviral immune responses. Recombinant baculovirus has been recently emerged as a new vaccine vector, since it has intrinsic immunostimulatory properties and good bio-safety profile.
METHODS
We have constructed a recombinant baculovirus-based RSV vaccine, Bac-RSV/G, displaying G glycoprotein, and evaluated immunogenicity and protective efficacy by intranasal immunization of BALB/c mice with Bac-RSV/G.
RESULTS
Bac-RSV/G efficiently provides protective immunity against RSV challenge. Strong serum IgG and mucosal IgA responses were induced by intranasal immunization with Bac-RSV/G. In addition to humoral immunity, G-specific Th17- as well as Th1-type T-cell responses were detected in the lungs of Bac-RSV/G-immune mice upon RSV challenge. Neither lung eosinophilia nor vaccine-induced weight loss was observed upon Bac-RSV/G immunization and subsequent RSV infection.
CONCLUSION
Our data demonstrate that intranasal administration of baculovirus-based Bac-RSV/G vaccine is efficient for the induction of protection against RSV and represents a promising prophylactic vaccination regimen.

Keyword

Respiratory syncytial virus; Glycoprotein; Recombinant baculovirus; Protective immunity

MeSH Terms

Administration, Intranasal
Animals
Baculoviridae
Eosinophilia
Glycoproteins
Immunity, Humoral
Immunization
Immunoglobulin A
Immunoglobulin G
Lung
Mice
Respiratory Syncytial Viruses
Respiratory Tract Diseases
T-Lymphocytes
Vaccination
Weight Loss
Glycoproteins
Immunoglobulin A
Immunoglobulin G

Figure

• 

  Figure 1 Construction and characterization of Bac-RSV/G vaccine. (A) The shuttle vector, pFastBac-RSV/G, was designed as shown in the diagram and constructed as described in the Materials and Methods. The vector was used to generate Bac-RSV/G recombinant baculovirus. (B) The presence of the RSV G protein on Bac-RSV/G was confirmed by western blotting. Viral particles were prepared as described in the Materials and Methods, and subjected to western blotting with an RSV G-specific monoclonal antibody.

• 

  Figure 2 Characterization of humoral immune responses induced by Bac-RSV/G vaccine. (A) BALB/c mice were immunized twice with 2×108 PFU of recombinant baculovirus (Bac-RSV/G or Bac-control) via intranasal route, and systemic anti-RSV IgG antibody titers were measured by serum ELISA two weeks after boosting. (B) Secretory IgA titers were measured in the BAL fluid for each immune group five days after RSV challenge. The results represent Log2 endpoint values from four to five individual mice. Data are representative of at least three independent experiments and data are average±SEM, n=4~5 mice per group. N.D., not detected.

• 

  Figure 3 G-specific Th1-cell response in Bac-RSV/G-immune mice. (A) Mice were i.n. immunized with Bac-RSV/G, Bac-control, or PBS and challenged with RSV A2 four weeks after boosting. Lung mononuclear cells were prepared from the lungs of the same group of mice (n=4~5) five days after challenge and stimulated with G183-195 peptide (+) or without peptide (-). Cells were stained for CD4, CD44, and IFN-γ, and analyzed by flow cytometry. Cells gated for CD4 are shown in each dot plot and the percentages represent the frequency of G-specific IFN-γ-positive cells. (B) Average data represent mean±SD (n=5). The results are a representative of two independent experiments.

• 

  Figure 4 G-specific Th17-cell response in Bac-RSV/G-immune mice. (A) Mice were immunized with Bac-RSV/G, Bac-control, or PBS i.n. and challenged with RSV four weeks after boosting. Lung mononuclear cells were prepared from the lungs of the same group of mice (n=4~5) five days after challenge and stimulated with G183-195 peptide (+) or without peptide (-). Cells were stained for CD4, CD43, and IL-17A, and analyzed by flow cytometry. Cells gated for CD4 are shown in each dot plot and the percentages represent the frequency of G-specific IL-17A-positive cells. (B) Average data represent mean±SD (n=5). The results are a representative of two independent experiments.

• 

  Figure 5 Bac-RSV/G immunization does not induce Th2 type cytokines in the lung. Mice were immunized and challenged as in Fig. 3. Lungs from mice were harvested five days after RSV challenge, and cytokine levels in the lung supernatant were assessed by multiplex antibody-based assay (FlowCytomix). Bars represent the mean of 4~5 mice per group and data are a representative of two independent experiments.

• 

  Figure 6 Immune protection from respiratory RSV challenge by vaccination with Bac-RSV/G. Each group of immune mice was challenged with 1×106 PFU of RSV A2 at 4 weeks after immunization and the level of viral replication in the lungs was determined by plaque assay at day 5. Results are expressed as the mean±SEM from 4 to 5 mice per group. The limit of detection is 200 PFU/g of lungs. N.D., not detected.

• 

  Figure 7 Bac-RSV/G immunization promotes neither lung eosinophilia nor vaccine-enhanced illness. (A) Mice were immunized and challenged as in Fig. 3. BAL was performed five days after RSV challenge, and BAL cells were stained with antibodies to CD45, Siglec-F, and CD11c, and eosinophils were quantitated among CD45+-gated cells. Average percentages of Siglec-F+CD11c-cells among total CD45+ cells represent mean±SD (n=6). The results are a representative of three independent experiments. (B) The same group of immune mice were challenged with RSV and then weighed each day. Results are expressed as the mean±SEM from 5 mice for each group. All data are representative of at least two independent experiments.

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